Copyright (c) 2025 C.A. Sri Ranjani, C. Asha Deepti
This work is licensed under a Creative Commons Attribution 4.0 International License.
Quantitative LC-MS/MS Determination of Source-Specific Variations of Underivatized Free Amino Acids in Protein Powders
Corresponding Author(s) : C.A. Sri Ranjani
Asian Journal of Chemistry,
Vol. 37 No. 7 (2025): Vol 37 Issue 7, 2025
Abstract
This study reports a novel, validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to comprehensively profile underivatized free amino acids (FAAs) in diverse whey and plant-based protein powders from global sources, which are commonly consumed for their nutritional value. Though their overall protein composition is frequently promoted, the accurate profiles and patterns of unmodified FAAs, whose functions are significant in metabolism, taste and product quality, are less well described. The validated LC-MS/MS technique showed a broad linearity range, high precision, accuracy and recoveries of 75-121%, with low detection and quantification limits. The analysis and separation time of less than 9 min increases its efficiency. Substantial differences in the FAAs present and their levels, both in kind and quantity, were observed between whey and plant-based proteins from various geographical region. The amino acids like glutamine and L-tryptophan were consistently high-ranking in most species of whey and plant-based proteins investigated. Notably, amino acids linked to sweet taste such as glycine, alanine, serine, proline and threonine were significantly greater in the majority of whey proteins that directly causes acute disorders in healthy individuals as per the International Council on Amino Acid Science (ICAAS). Principal component analysis (PCA) was capable of separating the protein samples that were tested which accounted for 86.88% of the total variance, thus verifying different clustering by FAA profiles. These observations reinforce the significance of source diversity and geographical origin in characterizing the nutritional profile of protein powders beyond bulk protein content. The large-scale profiling of underivatized FAAs furnishes rich information useful for consumers, manufacturers and researchers interested in deciphering and optimizing the nutritional, functional properties of these ubiquitous dietary supplements.
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D. Paddon-Jones and B.B. Rasmussen, Curr. Opin. Clin. Nutr. Metab. Care, 12, 86 (2009); https://doi.org/10.1097/MCO.0b013e32831cef8b
D.V. Martinho, H. Nobari, A. Faria, A. Field, D. Duarte and H. Sarmento, Nutrients, 14, 4002 (2022); https://doi.org/10.3390/nu14194002
J. Maughan, J. Sports Sci., 23, 883 (2005); https://doi.org/10.1080/02640410400023258
L. Keyes, A. Connaughton, M. O’Reilly, J. Walton and A. Flynn, Proc. Nutr. Soc., 71, 192 (2012); https://doi.org/10.1017/S0029665112002492
F. Bellisle, Am. J. Clin. Nutr., 99(Suppl 3), 738S (2014); https://doi.org/10.3945/ajcn.113.072210
G. Wu, Amino Acids, 37, 1 (2009); https://doi.org/10.1007/s00726-009-0269-0
P. Li, Y.L. Yin, D. Li, S.W. Kim and G. Wu, Amino Acids, 37, 229 (2007); https://doi.org/10.1017/S000711450769936X
A. Moran, S.L. Asa, K. Kovacs, E. Horvath, W. Singer, U. Sagman, J.-C. Reubi, C.B. Wilson, R. Larson and O.H. Pescovitz, N. Engl. J. Med., 323, 329 (1990); https://doi.org/10.1056/NEJM199008023230507
E.W. Boyer and M. Shannon, N. Engl. J. Med., 352, 1112 (2005); https://doi.org/10.1056/NEJMra041867
P.K. Gillman, J. Clin. Psychopharmacol., 25, 625 (2005); https://doi.org/10.1097/01.jcp.0000186741.30019.a3
L. You, C. Liu, H. Tang, Y. Liao and S. Fu,, Curr. Pharm. Des., 20, 3749 (2014); https://doi.org/10.2174/13816128113199990595
R. Dantzer, J.C. O’Connor, G.G. Freund, R.W. Johnson and K.W. Kelley, Nat. Rev. Neurosci., 9, 46 (2008); https://doi.org/10.1038/nrn2297
V. Bhatnagar, G. Xu, B.A. Hamilton, D.M. Truong, S.A. Eraly, W. Wu and S.K. Nigam, J. Hum. Genet., 51, 575 (2006); https://doi.org/10.1007/s10038-006-0398-1
B. O’Callaghan, A.M. Bosch and H. Houlden, J. Inherit. Metab. Dis., 42, 12 (2019); https://doi.org/10.1002/jimd.12053
P.E. Wischmeyer, Curr. Opin. Clin. Nutr. Metab. Care, 15, 195 (2012); https://doi.org/10.1097/MCO.0b013e32834f3b01
P. Newsholme, ImmunoTargets Ther., 2, 9 (2001); https://doi.org/10.2147/ITT.S44026
B. Socas-Rodríguez, J. González-Sálamo, J. Hernández-Borges and M.Á. Rodríguez-Delgado, Trends Analyt. Chem., 94, 148 (2017); https://doi.org/10.1016/j.trac.2017.07.002
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P. Chaimbault, K. Petritis, C. Elfakir and M. Dreux, J. Chromatogr. A, 855, 191 (1999); https://doi.org/10.1016/S0021-9673(99)00685-8
B. Buszewski and S. Noga, Anal. Bioanal. Chem., 402, 231 (2012); https://doi.org/10.1007/s00216-011-5308-5
H.G. Gika, G.A. Theodoridis, U. Vrhovsek and F. Mattivi, J. Chromatogr. A, 1259, 121 (2012); https://doi.org/10.1016/j.chroma.2012.02.010
B. Álvarez-Sánchez, F. Priego-Capote and M. Castro, Trends Analyt. Chem., 29, 120 (2010); https://doi.org/10.1016/j.trac.2009.12.004
Y. Takano, Y. Kashiyama, N.O. Ogawa, Y. Chikaraishi and N. Ohkouchi, Rapid Commun. Mass Spectrom., 24, 2317 (2010); https://doi.org/10.1002/rcm.4651
L. Wang, R. Xu, B. Hu, W. Li, Y. Sun, Y. Tu and X. Zeng, Food Chem., 123, 1259 (2010); https://doi.org/10.1016/j.foodchem.2010.05.063
B.M. Silva, S. Casal, P.B. Andrade, R.M. Seabra, M.B. Oliveira and M.A. Ferreira, Anal. Sci., 19, 1285 (2003); https://doi.org/10.2116/analsci.19.1285
V. Fierabracci, P. Masiello, M. Novelli and E. Bergamini, J. Chromatogr., Biomed. Appl., 570, 285 (1991); https://doi.org/10.1016/0378-4347(91)80531-G
M. Alaiz, J.L. Navarro, J. Girón and E. Vioque, J. Chromatogr. A, 591, 181 (1992); https://doi.org/10.1016/0021-9673(92)80236-N
C. Megías, I. Cortés-Giraldo, J. Girón-Calle, J. Vioque and M. Alaiz, Talanta, 131, 95 (2015); https://doi.org/10.1016/j.talanta.2014.07.077
S.A. Cohen and K.M. De Antonis, J. Chromatogr. A, 661, 25 (1994); https://doi.org/10.1016/0021-9673(93)E0821-B
J. Boye and C. Barbana, Protein Processing in Food and Bio-product Manufacturing and Techniques of Analysis, In: Food and Industrial Bioproducts and Bioprocessing, Wiley-Blackwell: New York, Chap. 3, pp. 85-114 (2012).
G. Joana Gil-Chávez, J.A. Villa, J. Fernando Ayala-Zavala, J. Basilio Heredia, D. Sepulveda, E.M. Yahia and G.A. González-Aguilar, Compr. Rev. Food Sci. Food Saf., 12, 5 (2013); https://doi.org/10.1111/1541-4337.12005
S. Pedersen-Bjergaard, K.E. Rasmussen and T. Grønhaug Halvorsen, J. Chromatogr. A, 902, 91 (2000); https://doi.org/10.1016/S0021-9673(00)00738-X
